Stem Cell Genetics Laboratory

Since its foundation, the Laboratory has worked on development of new therapeutic approaches for reparative osteogenesis using tissue engineering constructions (TEC) based on osteoprogenator cells and osteoplastic materials, which may include protein factors or gene constructs with osteoinductive action. A cryobank of cell cultures of multipotent mesenchymal stromal cells (MMSCs) obtained from various sources — bone marrow, adipose tissue, umbilical cord vessels, tooth pulp of humans and animals.

The effect of bone inducers such as dexamethasone, dihydroxyvitamin D3 and bone morphogenetic protein BMP-2 on MMSC cultures was studied. The efficacy of using TECs based on MMSCs that were pre-differentiated in the osteogenic direction is shown in vivo in models of heterotopic osteogenesis and during transplantation into the area of bone defects. MMSC-based TEC from a pulp of fallen out milk teeth has been successfully tested for pulp tissue regeneration.

Osteoplastic materials based on polymers of natural and synthetic origin - collagen, chitosan, polylactic and polyglycolic acids, in the form of porous matrices obtained by foaming, laser sintering, 3D printing, and in the form of thermosetting hydrogels are used as cells matrices. Methods of obtaining the materials provide the possibility of impregnation of biologically active substances, which effect is aimed at reducing inflammatory reactions during implantation, enhancing vascularization and osteoinductive action in order to increase the regenerative potential of TEC. The use of non-steroidal anti-inflammatory ibuprofen as part of polylactide matrices significantly increases cell survival during allogeneic transplantation into the bone defect area, ensuring the development of direct osteogenesis without cartilage tissue formation (indirect osteogenesis) and as a result, significantly accelerates the repair and restoration of functionally active bone tissue. TEC uses platelet-rich plasma - a rich source of growth factors, such as FGF, VEGF, PDGF, EGF, and others, providing neovasculogenesis inside the TEC and attracting resident predecessors.

A method of gene therapy ex vivo was developed. MMSCs of adipose tissue were transduced with adenoviral constructs carrying BMP-2 gene. Transplantation of tissue engineering constructions with transduced cells leads to long-term production of a target protein, which ensures a high therapeutic effect of such product. The role of non-coding RNAs in the regulation of osteogenesis is analyzed to increase the efficiency of osteogenic differentiation. New gene products for reparative osteogenesis are being developed using transient viral constructs and plasmid vectors with high transfection efficiency.

In 2018, the Stem Cell Genetics Laboratory introduced a method for obtaining organelles stable cultures from human intestinal biopsies. Intestinal organelles imitate the structure of intestinal epithelium, and formed by several cell types and therefore they are miniaturized and simplified versions of the organ in vitro. The use of organoids provides the opportunity to treat many diseases, to create and test new drugs. The Laboratory implements and optimizes medical technology for the use of intestinal organoids for the selection of personalized cystic fibrosis therapy, which diagnosis and treatment significantly complicates the allelic heterogeneity of CFTR. The in vitro test bases on obtaining organoids from a patient's intestinal biopsy material and treating them with forskolin. The effectiveness of targeted drugs in patients with cystic fibrosis in Russia, including those with rare and unique mutations of CFTR gene, will be assessed using this approach. A cryobank with intestinal organoids was created, which is replenished with new unique cultures. The Laboratory collaborates with the forskolin test developers from the University Medical Center Utrecht in the Netherlands (Molecular cystic fibrosis laboratory of the University Medical Center Utrecht).

The Stem Cell Genetics Laboratory is implementing a project on developing a new nootropic drug based on juvenile neuropeptides derived from the human induced pluripotent stem cells (IPSC). Unlike the currently existing drugs from xenogenic animal tissues, this medication will contain allogeneic or autologous components, which will ensure its low immunogenicity while retaining full-length peptides with pronounced functional activity

The Stem Cell Genetics Laboratory participates in the implementation of multidisciplinary research projects supported by the Russian Science Scientific Foundation and the Russian Federation Ministry of Science grants, in collaboration with research teams from the Kurchatov Institute National Research Center, Central Research Institute of Dentistry and Oral and Maxillofacial Surgery under the Ministry of Health, the Federal Research Center for Crystallography and Photonics RAS, Scientific Research Institute of Human Morphology and others.

Four theses for candidate degree were defended based on the research materials of the Laboratory.

1. Bukharova T.B., Antonov E.N., Popov V.K., Fatkhudinov T.Kh., Popova A.V., Volkov A.V., Bochkova S.A., Bagratashvili V.N., Goldshtein D.V. Biocompatibility of tissue engineering constructions from porous polylactide carriers obtained by the method of selective laser sintering and bone marrow-derived multipotent stromal cells. Bull Exp Biol Med. 2010 Jul; 149(1):148-53.
2. Bukharova T.B., Arutyunyan I.V., Shustrov S.A., Alekseeva I.S., Fedyunina I.A., Logovskaya L.V., Volkov A.V., Rzhaninova A.A., Grigoryan A.S., Kulakov A.A., Goldshtein D.V. Tissue engineering construct on the basis of multipotent stromal adipose tissue cells and Osteomatrix for regeneration of the bone tissue. Bull Exp Biol Med. 2011 Nov;152(1):153-8.
3. Logovskaya L.V., Bukharova T.B., Volkov A.V., Vikhrova E.B., Makhnach O.V., Goldshtein D.V. Induction of osteogenic differentiation of multipotent mesenchymal stromal cells from human adipose tissue. Bull Exp Biol Med. 2013 May;155(1):145-50.
4. Bukharova T.B., Logovskaya L.V., Volkov A.V., Garas M.N., Vikhrova E.B., Logunov D.Y., Makhnach O.V., Shmarov M.M., Goldshtein D.V. Adenoviral transduction of multipotent mesenchymal stromal cells from human adipose tissue with bone morphogenetic protein BMP-2 gene. Bull Exp Biol Med. 2013 Nov;156(1):122-6.
5. Bukharova T.B., Leonov G.E., Galitsyna E.V., Vasilyev, A.V., Vakhrushev I.V., Vikhrova E.B., Makhnach O.V., Goldstein D.V. Osteogenic potential of multipotent mesenchymal stromal cells from human exfoliated deciduous teeth before and after cryopreservation. Genes and Cells. 2016. 11(4), с. 43-47.
6. E. N. Antonov, T. B. Bukharova, A. G. Dunaev, L. I. Krotova, I. E. Nifant’ev, V. K. Popov, A. V. Shlyakhtin. New “old” polylactides for tissue engineering constructions. Inorganic Materials: Applied Research, 2017, Vol. 8, No. 5, pp. 704–712.
7. Kulakov A.A., Goldshtein D.V., Krechina E.K., Bukharova T.B., Volkov A.V., Gadzhiev A.K. Regeneration of dental pulp tissue using pulpal autologous mesenchymal stem cells and platelet-rich plasma. Stomatologiia (Mosk). 2017; 96(6):12-16.
8. Grigoriev T.E., Zagoskin Y.D., Belousov S.I., Vasilyev A.V., Bukharova T.B., Leonov G.E., Galitsyna E.V., Goldshtein D.V., Chvalun S.N., Kulakov A.A., Paltsev M.A. «Influence of molecular characteristics of chitosan on properties of in situ formed scaffolds». BioNanoScience. 2017. 7(3). pp. 492-495.
9. T.E. Grigoriev, T.B. Bukharova, A.V. Vasilyev, G.E. Leonov, Y.D. Zagoskin, V.S. Kuznetsova, V. I. Gomzyak, D.I. Salikhova, E. V. Galitsyna, O.V. Makhnach, K. V. Tokaev, S. N. Chvalun, D.V. Goldshtein, A.A. Kulakov, M A. Paltsev. Effect of molecular characteristics and morphology on mechanical performance and biocompatibility of PLA-based spongious scaffolds. «BioNanoScience» (2018).
10. Efremova A.S., Bukharova T.B., Kashirskaya N.Yu., Goldstein D.V. The use of intestinal organelles for personalized diagnosis and treatment of cystic fibrosis. Medical Genetics. 2018; Vol. 17, No. 9 (194), p. 3-12